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Article

Characterization of Volatile Compounds in Traditional Mexican Cocoa-Based Beverages and Their Application in Flavored Milks

by
Lucía Abadía-García
1,
Edgar Ayala-Padilla
1,
Pedro A. Vázquez-Landaverde
2,
María del Carmen Cortez-Trejo
1 and
Silvia L. Amaya-Llano
1,*
1
School of Chemistry, Autonomous University of Querétaro, Santiago de Querétaro 76010, Querétaro, Mexico
2
Center for Research in Applied Science and Advanced Technology, National Polytechnic Institute, Querétaro Unit, Santiago de Querétaro 76090, Querétaro, Mexico
*
Author to whom correspondence should be addressed.
Beverages 2025, 11(5), 130; https://doi.org/10.3390/beverages11050130
Submission received: 19 June 2025 / Revised: 16 August 2025 / Accepted: 27 August 2025 / Published: 2 September 2025
(This article belongs to the Special Issue New Insights into Artisanal and Traditional Beverages)
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Abstract

Traditional Mexican non-alcoholic, cocoa-based beverages are a relevant part of the gastronomy of the country. This study aimed to identify the volatile compound profile of three traditional beverages from Mexican gastronomy, including Pozol, Chilate, and Cacahuatole, and evaluate the acceptability of milks incorporating their traditional flavors. Chemical composition was determined, and volatile compounds were analyzed using gas chromatography–mass spectrometry (GC-MS). The results showed the Mexican beverages are rich in carbohydrates (70.3–78.2% d.b.), proteins (9.3–16% d.b.), and fat (8.8–11.7% d.b.) due to corn, rice, broad beans, and cocoa ingredients. GC revealed volatile profiles, with Pozol containing 148, Chilate 104, and Cacahuatole 109 compounds, mainly terpenes, pyrazines, aldehydes, and phenolics. Nixtamalization, cocoa roasting, and ingredient variations influenced the volatile composition and sensory attributes. Multivariate analysis distinguished the beverages based on their volatile composition, revealing ingredient-dependent variations. Artificial flavors replicating these profiles were developed and incorporated into milks, which were evaluated by a trained sensory panel using the Check-All-That-Apply (CATA) method. Sensory analysis confirmed that key aromatic compounds contributed to flavor perception. This work identified for the first time the volatile compounds of traditional Mexican cocoa-based beverages, namely, Pozol, Chilate, and Cacahuatole, and provides insight into the role of traditional preparation methods in the development of flavor profiles. On the other hand, this study highlights the potential for incorporating traditional Mexican beverage flavors into commercial milk for novel product development.

Graphical Abstract

1. Introduction

Traditional Mexican cuisine is recognized as an essential part of the world’s intangible cultural heritage, designated by the United Nations Educational, Scientific, and Cultural Organization (UNESCO) in 2010 [1]. Among its culinary traditions, Mexico possesses a rich variety of cocoa-based beverages, many of which date from pre- and post-Hispanic times, are derived from indigenous traditions, and have been passed down through generations. Cocoa-based beverages represent a significant element of traditional Mexican gastronomic history. These traditional drinks, including Pozol, Cacahuatole, and Chilate, are commonly consumed in different regions of the country and are distinguished by their unique flavor profiles, which result from complex combinations of cocoa with cereals and spices, and specific preparation techniques such as nixtamalization, drying, baking, and roasting processes [2,3].
Pozol is one of the most ancient endemic Mesoamerican products still consumed in Mexico [1]. It is a corn-based beverage with cocoa, cinnamon, and other ingredients added to produce a nonstick dough that is diluted in water, and sugar is added before consumption. It is a typical beverage for the Mayan region, including Chiapas, Tabasco, Campeche, and Yucatan states. Cacahuatole, on the other hand, is a drink made from cocoa, corn, and broad beans and is mainly prepared in central Mexico, mostly in Tlaxcala. It is elaborated in the form of a paste, which must also be diluted before consumption [4]. Chilate is produced in Guerrero; the elaboration of this beverage includes cocoa, sugar, cinnamon, and rice, and it can be diluted in water or milk [5,6].
Despite their cultural and historical significance, there is a notable lack of scientific literature regarding the characterization of non-alcoholic traditional Mexican cocoa-based beverages. While previous studies have explored the sensory and nutritional properties of cocoa and corn-based foods [7,8], little research has been conducted on the volatile compound profiles that define the aroma and flavor of these traditional drinks. Aroma is a crucial factor in food acceptability, and the understanding of the volatile components of these beverages can contribute to preserving their authenticity while exploring their potential applications in food science and product development [9,10].
In addition to their cultural importance, cocoa-based beverages offer an opportunity for innovation in the dairy industry. Milk consumption in Mexico remains below the recommended intake levels set by the Food and Agriculture Organization (FAO), particularly among lower-income populations [11]. Given the affordability and nutritional potential of milk made from milk solids and vegetable fats, they present a viable means of addressing dietary gaps [12,13]. Since chocolate is one of the most widely preferred flavors in dairy beverages [14], integrating the flavor profiles of traditional Mexican cocoa-based beverages into milks could enhance their acceptability and consumption.
This work aimed to identify the volatile compound profile of three traditional cocoa-based beverages from Mexican gastronomy, namely, Pozol, Chilate, and Cacahuatole, and evaluate the acceptability of milks with their traditional flavors. Samples of the beverages were collected at local sites in different Mexican states while verbally interviewing master artisans about the fabrication process, and the volatile compounds present in the beverages were identified using gas chromatography–mass spectrometry (GC-MS) analysis. The volatile compound profiles provided valuable insights for the dairy industry, particularly allowing for the development of flavored milk-based beverages oriented to the improvement of the diet of people in precarious socioeconomic situations in Mexico; the sensory evaluation of the milks with the flavors of the traditional Mexican beverages was assessed using a semi-trained consumer panel to evaluate their acceptability. This study contributes to the preservation and valorization of Mexican cocoa-based beverages, bridging the gap between traditional culinary heritage and contemporary food science.

2. Materials and Methods

2.1. Materials and Chemicals

In total, 3–5 samples of paste/dough for traditional beverages were obtained from local markets: Pozol from Tuxtla Gutiérrez (Chiapas, Mexico), Cacahuatole from Zacatelco (Tlaxcala, Mexico), and Chilate from Acapulco (Guerrero, Mexico). Interviews were conducted with the artisans to record the beverage production processes. Milk (Fortileche Plus®, Alpura®, Cuautitlán Izcalli, Mexico) and sugar were purchased from a local store, and all HPLC-grade and reagent-grade chemicals were acquired from Sigma-Aldrich (Saint Louis, MO, USA) and JT Baker (City of Mexico, State of México, Mexico) respectively.

2.2. Chemical Composition of Pastes/Doughs

The chemical composition of Pozol, Cacahuatole, and Chilate pastes/doughs was determined according to AOAC methods [15] for moisture (930.15), fat (7.056), protein (942.05), ash (942.05), and crude fiber (973.19). Carbohydrate content was calculated on dry weight basis as follows: 100-(protein content + ashes + fat + fiber). Metabolizable energy was calculated using the Atwater conversion factors for whole cornmeal: protein 11.4 kJ/g (2.73 kcal/g), fat 35 kJ/g (8.37 kcal/g), and carbohydrate 16.9 kJ/g (4.03 kcal/g) [16].

2.3. Identification of Volatile Compounds

The volatile compound profile of the pastes/doughs was determined according to [17]. Briefly, 3 g of the samples were hermetically sealed into a 20 mL vial with an EFRP/silicone stopper and stored frozen at −22 °C until analysis. Analysis of volatile compounds was carried out using a gas chromatograph (Agilent 7890A, Santa Clara, CA, USA) coupled with an ionizing flame detector (FID), a mass spectrometer (Agilent 5975C), and an HP-5MS column (60 m × 0.25 nm ID × 0.25 μm DF, Agilent, Santa Clara, CA, USA) using Helium as a carrier gas (flow rate, of 1 mL/min; pressure, 124 kPa). Before analysis, the vials were conditioned at 50 °C for 5 min, and a 2 cm 50/30 μm divinylbenzene/carboxene/polydimethylsiloxane (DVB/CAR/PDMS) fiber (Supelco, Bellefonte, PA, USA) was exposed for 60 min at 50 °C to the headspace of the thawed aliquot in the vial for the adsorption of the analytes. The fiber was desorbed for 10 min at the injection point, and the sample was injected at 200 °C in splitless mode. The oven temperature was set at 60 °C for the first 5 min and was increased until reaching 150 °C (rate 4 °C/min), maintaining this condition for 20 min. Then, the temperature was raised to 220 °C at a rate of 3 °C/min and was held for 5 min. A final increase was made at a rate of 6 °C/min until reaching 240 °C, maintaining this condition for 5 min. The volatile compounds were detected using a temperature of 230 °C and 150 °C for the source and quadrupole, respectively. The mass spectra were acquired in a scanning mode in a range of 33–600 m/z. The equipment was operated in electron impact ionization mode at 70 eV. Data analysis was carried out using the MSD ChemStation software E.01.00.237 (Agilent Technologies Inc., Santa Clara, CA, USA). The compound identification was made by comparing the mass spectra with those found in the National Institute of Standards and Technology (NIST, Gaithersburg, MD, USA) 08 library, where the mass spectra found were compared with those available in the literature [18,19].

2.4. Flavor Development Based on Mexican Beverage Volatile Compound Profile

To replicate the characteristic sensory attributes of the traditional Mexican beverages (Pozol, Cacahuatole, and Chilate), collaborations were established with four commercial flavor manufacturers: Takasago International Co. (Tlalnepantla, State of México, Mexico), DSM Firmenich (México City, Mexico), International Flavors & Fragrances Inc. (Tlalnepantla, State of México, Mexico), and MANE Flavor & Fragrance Manufacturer (Toluca, State of México, Mexico). Each company was provided with detailed compositional data, including the ingredient lists and the volatile compound profiles previously determined for each beverage. Based on this information, the companies formulated artificial flavor prototypes designed to mimic the beverage profiles. The resulting flavor samples were supplied as liquid concentrate and applied depending on the manufacturer’s production specifications.

2.5. Flavored Milk Fabrication

The flavors were added to commercial milk (Fortileche Plus®, Alpura®, Cuautitlán Izcalli, Mexico), and the percentage of application was made according to the doses recommended by the flavorist companies. Traditional beverage controls made with Pozol, Chilate, and Cacahuatole dough combined with milk and sugar were used (Supplementary Material, Table S1).

2.6. Sensory Evaluation of Flavored Milks

The flavored milks were evaluated by a trained sensory panel composed of students from the Autonomous University of Querétaro. In previous sessions, the panel consumed the traditional drinks, one beverage per week, to identify their flavor. They were asked to describe traditional beverages about taste, smell, texture, and appearance. Then, 8–9 descriptive terms of flavor were generated (Supplementary Material, Table S2). Also, descriptive phrases specific to each beverage were established when participants were familiar with the sensorial characteristics of traditional beverages following the “free choice profiling” method [20] (Supplementary Material, Table S3). In the sensory evaluation, the panel evaluated the flavor proposals for each sample according to ISO Standard 6658:2017 [21]. Flavor proposals were given together with a control of traditional drinks prepared with milk added with sucrose. A multivariate Check All That Apply (CATA) method was applied. For this purpose, the predetermined list of phrases describing each beverage was presented, and the evaluators were asked to select the best phrases that represent the sensory attributes of milks. The samples were coded with four random numbers and presented at 10 °C in a 25 mL portion. Water and crackers were offered to cleanse the palate between each sample. The sensory evaluation protocol was approved by the Bioethics Committee of the Faculty of Chemistry (Autonomous University of Querétaro).

2.7. Statistical Analysis

At least 3 samples of traditional Mexican beverages were evaluated. The volatile compound profiles were coded binary, with 0 and 1, for the absence and presence of a specific compound, respectively. These encoded profiles were analyzed with a Venn diagram using an online tool [Bioinformatics and Systems Biology of Gent, URL: http://bioinformatics.psb.ugent.be/webtools/Venn/ (accessed on 13 August 2025)]. Then, multivariate analysis such as principal component analysis and dendrogram were carried out using the percentage of area under the chromatogram curve with R Studio version 4.4.2 and JMP 8.0 software (SAS Institute Inc., Cary, NC, USA), respectively. For CATA analysis, R Studio was used.

3. Results and Discussion

3.1. Composition and Preparation Methods

The proximate composition of pastes/doughs of the traditional beverages is presented in Table 1. Among the components examined, moisture content exhibited the greatest variation, likely due to differences in processing methods. The nixtamalization process, used in the preparation of Pozol and Cacahuatole, plays a crucial role in these variations. Nixtamalization is a traditional technique that transforms corn kernels into a cohesive dough called “masa” by cooking them in an alkaline solution with calcium hydroxide. This process induces significant physicochemical changes, facilitating starch gelatinization and its interaction with calcium, which influences pasting properties, digestibility, and hydration characteristics [22]. Consequently, nixtamalization increases moisture content, contributing to the formation of a more elastic and cohesive dough.
In contrast, Chilate exhibited a lower moisture content than Pozol and Cacahuatole, a difference attributed to its processing method. The preparation of Chilate involves soaking rice for approximately two hours, allowing for gradual water absorption. However, moisture uptake during rice soaking is influenced by factors such as temperature, initial moisture content, and rice variety. Notably, corn starch generally has a higher water absorption capacity than rice starch, resulting in lower moisture content in Chilate [23]. These findings highlight the critical role of processing techniques in shaping the hydration mechanisms of starch, which in turn modify the composition and texture of traditional Mexican beverages.
The protein and lipid content of the beverages is primarily influenced by the amount of cocoa beans used in their preparation. Cocoa beans are notable for their high lipid and protein content, with fat comprising between 40 to 57% of their composition. Their fatty acid profile consists primarily of palmitic, stearic, and oleic acids. Notably, oleic acid has recognized benefits for cardiovascular health; thus, its presence alongside saturated fatty acids may help mitigate adverse effects and present advantageous opportunities for the food industry [24]. Protein concentration in cocoa beans ranges from 11.8 to 15.7% on a dry-weight basis, higher than that of corn, which varies from approximately 9.2 to 15.8%. Cocoa proteins primarily consist of albumins (52%) and globulins (43%), which play essential roles in the structural and functional properties of the beans, influencing their nutritional profile and behavior during processing [25].
The three analyzed beverage recipes include cocoa beans as a key ingredient; cocoa beans are the most expensive component and contribute to variability among local producers. Consequently, slight differences in fat and protein content across samples can be attributed to variations in cocoa bean inclusion. Similar findings were reported by [26]. The study analyzed Pozol dough samples from Tabasco, México, and found comparable compositions. Cacahuatole dough exhibited higher protein content due to the incorporation of broad beans. Broad beans have a high protein content, typically between 27 and 34% on a dry matter basis, with globulins and albumins as the primary protein fractions [27]. Their high-quality protein profile, characterized by an adequate quantity of essential amino acids, enhances the nutritional value of Cacahuatole.
The fiber content of the beverages ranged from 2.0 to 2.4%, similar to values reported in Tejate samples by Sotelo et al. [28]. This confirms that these beverages are low in fiber. The dehulling of cocoa and broad beans after roasting contributes to this low fiber content. A notable advantage of dehulling and toasting these beans is the reduction of antinutritional factors [27].
The caloric content of the doughs ranged from 1727 to 1763 kJ/100 g DW (412–420 kcal), similar to values reported by Gonzalez-Amaro [29] for Tejate dough, a comparable Mexican beverage rich in carbohydrates and fat derived from cacao and nixtamalized corn. However, these authors found that such beverages promote better glucose metabolism compared to juices and soft drinks. Specifically, significantly lower postprandial glycemic responses were observed, attributed to the formation of starch–lipid complexes in cacao–corn mixtures. These complexes, classified as type V resistant starch, are digested slowly by gastric enzymes and may undergo fermentation in the colon, offering potential health benefits to consumers.

3.2. Volatile Compounds

The preparation methods of the studied traditional Mexican beverages are presented in Figure 1; these processes involve various artisanal processes. Crucial steps such as alkaline cooking of corn (nixtamalization), rice soaking, roasting of cocoa beans, and stone grinding have been applied since pre-Hispanic times, contributing to the release and development of volatile compounds from their ingredients. The volatile compounds in the three beverages were identified using gas chromatography. Specifically, 145, 90, and 106 different compounds were detected in samples of Pozol, Chilate, and Cacahuatole, respectively (Supplementary Material, Table S4). In Pozol, the most abundant volatile groups included terpenes (31), alcohols (14), aldehydes (13), benzenes (10), pyrazines (9), and acids (6). Less abundant groups comprised alkanes, furans, ketones, and esters. Similarly, Chilate samples contained 34 terpenes, followed by aldehydes, alcohols, and acids, each with 7 to 13 compounds. Cacahuatole also exhibited a high number of terpenes (38), along with benzenes, aldehydes, pyrazines, and alcohols, each represented by 9–18 compounds. However, not all volatiles were consistently found across samples, indicating high variability in preparation methods and ingredients. Venn diagrams (Figure 2) revealed only 12 common compounds in Pozol, 15 in Chilate, and 27 in Cacahuatole. Table 2 summarizes these common volatiles found among replicates.
Due to the great diversity of identified volatiles, their analysis can only be interpreted from a general point of view and is mainly related to the ingredients and processing methods used. Across all beverages, terpenes dominated the volatile profiles (Table S4 and Table 2). These compounds impart fruity, floral, spicy, and citrus-like aromas. Major contributors included (-)-β-pinene, (-)-terpinen-4-ol, 1(e)-β-ocimene, α-caryophyllene, α-phellandrene, α-thujene, β-phellandrene, camphene, caryophyllene, γ-terpinene, and sabinene, which originate mainly from cocoa beans, cereals such as corn, rice [30,31], and legumes like broad beans [32].
Focusing on used ingredients, cocoa features a range of volatile compounds, derived from its inherent components as well as those formed during roasting. In this way, cocoa-derived volatiles include aldehydes such as 2-methyl propanal, 2-methyl butanal, and 3-methyl butanal, and ketones such as 2-heptanone, 2-pentanone, 2-nonanone, acetophenone, and acetoin, imparting chocolate and malty notes and enhancing cocoa flavor [33,34,35]. In this work, 3-methylbutanal (in PZ2 and CA2), 2-methylbutanal (in PZ2, CH2, CH5, and CA2), and 2-nonanone (in CH5) were detected (Table S4). In addition, cocoa roasting was a key source of pyrazines responsible for nutty, roasted aromas, with tetramethylpyrazine accounting for up to 90% of the total [34]. This pyrazine appeared in Chilate samples, while 2,3-dimethylpyrazine, 2,3-diethyl-5-methylpyrazine, 2,5-dimethylpyrazine, 3,5-diethyl-2-methylpyrazine, ethylpyrazine, and methylpyrazine were detected in Cacahuatole (Table 2). Concerning Pozol, several pyrazines were identified (e.g., methylpyrazine, ethylpyrazine, tetramethylpyrazine, etc.) (Table S4); however, they were not common to all the samples evaluated. Overall, results suggest differences in roasting intensity or cocoa bean quality among beverages [34].
Roasting processes involved in beverage production also promoted the formation of floral, fruity, and sweet esters and alcohols, including 1-butanol-3-methyl-acetate, 2-propen-1-ol-3-phenyl-acetate, 3-phenyl-2-propen-1-ol, and linalool (Table 2). Linalool is a key floral note in South American cocoa [35]; thus, its natural presence in cocoa beans may also be possible and would explain their detection in all the analyzed samples. Other detected volatiles, such as 2-phenylethyl acetate, 2-heptanol, hexanal, nonanal, benzeneacetaldehyde, and furfural, enriched the aroma [34,35,36].
Table 2. Volatile compounds a in Pozol, Chilate, and Cacahuatole.
Table 2. Volatile compounds a in Pozol, Chilate, and Cacahuatole.
CompoundCASAroma Descriptor bPZCHCACompoundCAS Aroma Descriptor b PZ CH CA
Terpenes:      Pyrazines:
(-)-Beta-Pinene000127-91-3Pine, woodXX-     Pyrazine, 2,3-dimethyl005910-89-4Caramel, cocoa, hazelnut, peanut butter, toasted--X
(-)-Terpinen-4-ol020126-76-5---X     Pyrazine, 2,3-diethyl-5-methyl018138-04-0Earthy, meat, potato, roasted--X
1(E)-Beta-Ocimene003779-61-1Floral-X-     Pyrazine, 2,5-dimethyl000123-32-0Cocoa, roast beef, toasted walnut --X
Alpha-Caryophyllene006753-98-6Fried, spices, wood-X-     Pyrazine, 3,5-diethyl-2-methyl018138-05-1Baked, cocoa, toasted, rum--X
Alpha-Phellandrene000099-83-2Citrus, fresh, mint, pepper, spices, and wood-XX     Pyrazine, ethyl013925-00-3Burnt, must, peanut butter, toasted, rum, wood--X
Alpha-Thujene002867-05-2--XX     Pyrazine, methyl000109-08-0Cocoa, green, hazelnut, popcorn, roasted--X
Beta-Phellandrene000555-10-2---X     Pyrazine, tetramethyl001124-11-4Cocoa, coffee, green, mocha, roasted-X-
Camphene000079-92-5Menthol, citrus-XX
Caryophyllene000087-44-5Fried, spices, woodX-X      Esters:
Gamma-Terpinene000099-85-4Bitter and citrusy-XX     1-Butanol, 3-methyl-, acetate000123-92-2Apple, banana, glue, pearX--
Sabinene003387-41-5---X     2-Propen-1-ol, 3-phenyl-, acetate000103-54-8Floral, fruit, honey--X
Acids:      Aldehydes:
Dodecanoic Acid/Lauric Acid000143-07-7-X--     Benzaldehyde000100-52-7Bitter almond-XX
Nonanoic Acid/Pelargonic Acid000112-05-0Fat and bitterX--     Benzene acetaldehyde000122-78-1 --X
     Cinnamaldehyde, (E)014371-10-9 --X
Alcohols:      Benzene derivatives:
2-Propen-1-ol, 3-Phenyl000104-54-1-X-X     Benzene, 1-methoxy-4-(1-
propenyl)		000104-46-1---X
Ethanol000064-17-5-XXX     Benzene, 1-methyl-4-(1-
methylethyl)		000099-87-6Fresh, citrusy-X-
Linalool000078-70-6Coriander, floral, lavender, lemon, roseXXX     Estragole 000140-67-0---X
     Phenylethyl alcohol000060-12-8Fruit, honey, lilac, rose, wine-X-
Phenols:      Others:
2-Methoxy-4-Vinylphenol007786-61-0Clove, curry, spicesX-X     2-Furanmethanol000098-00-0Burnt, caramel, cooked--X
Eugenol000097-53-0Burnt, cloves, spicesX-X     Hexane000110-54-3-XX-
     Styrene000100-42-5Sweet and floralXXX
a Compounds found in all samples of each beverage. b Most frequently associated aroma descriptors [37]. PZ, Pozol. CH, Chilate. CA, Cacahuatole.
Beyond flavor, cocoa roasting reduces astringency and polyphenol content, producing melanoidins that contribute to bean darkening. A balanced polyphenol retention is desirable to preserve antioxidant capacity while preventing flavor loss [34]. Detected phenols included 2-methoxy-4-vinylphenol (clove, curry, spicy notes) and eugenol (burnt, clove, spicy notes) in Cacahuatole and Pozol beverages (Table 2).
All the drinks analyzed contained cocoa, but their distinct ingredients led to unique volatile profiles. Pozol, which is made from cocoa and nixtamalized corn, differed noticeably from Chilate (featuring cocoa and rice) and Cacahuatole (which includes cocoa, corn, and broad beans), creating clear differences among them. For instance, the use of nixtamalized maize in the preparation of Pozol and Cacahuatole was anticipated to confer relevant volatile compounds. Nixtamalization is known to hydrolyze bound phytochemicals, enhancing their extractability [38]. Lipid-oxidation aldehydes such as hexanal, heptanal, octanal, and nonanal are common in nixtamalized corn [39]. These compounds are also related to broad beans [31,36]. Nevertheless, hexanal and nonanal were only detected in CA1; however, their absence in the other Cacahuatole samples and Pozol may be related to subsequent thermal decomposition. The use of broad beans in Cacahuatole, on the other hand, could be responsible for the detection of a greater amount of pyrazines, compared to Pozol and Chilate, since pyrazines were predominant in roasted broad beans [31].
Spices played a role in enhancing the beverage’s aroma. Analytical results indicated the presence of cinnamaldehyde, eugenol, and benzaldehyde from cinnamon in multiple samples. Cinnamaldehyde was found in Pozol (PZ2, PZ3, PZ5), Chilate (CH1, CH3), and Cacahuatole (CA1-3). In addition, eugenol was found in all samples of Pozol and Cacahuatole as well as CH1,2,5, and benzaldehyde was found in all samples of Chilate and Cacahuatole as well as PZ2-4. Furthermore, the detection of styrene in all three beverages (Table 2) can be attributed to the presence of cinnamon, which is known to naturally contain elevated levels of styrene [40,41] and may also generate it through the degradation of cinnamaldehyde [42]. Estragole, on the other hand, was found only in Cacahuatole samples (Table 2) and is chiefly associated with the anise seeds used in its preparation [43].
Although the pastes/doughs of the traditional Mexican beverages are not deliberately fermented, ethanol was consistently found in all beverage samples (Table 2), likely due to the activity of lactic acid bacteria (LAB) during storage at ambient temperature between preparation and consumption [2,44]. Additional cocoa fermentation-associated compounds, including guaiacol and terpineol [45], were also detected in specific samples. Guaiacol was present in PZ2, while terpineol was identified in PZ2, CH1-4, and CA3.
The results of this study provided an initial assessment of the volatiles identified in traditional Mexican beverages. Quantifying these concentrations was not part of the current analysis. Future research should include quantification, as the sensory relevance of volatiles is determined by both their presence and concentration, as well as interactions with other compounds within the beverage matrix. The relationship between volatile compound concentrations and their odor thresholds plays a key role in understanding aroma perception and overall acceptability. The olfactory threshold refers to the lowest concentration at which a particular compound can be detected by the human nose. Volatile compounds with concentrations above this threshold contribute to aroma, while those below the threshold may still affect perception through interactions with other volatiles. Some key volatile compounds in cocoa have well-established olfactory thresholds, which are used by the chocolate industry to monitor and control flavor quality. Compounds such as pyrazines, esters, acids, and alcohols have thresholds in the low microgram per kilogram range. A few threshold values for some of the identified compounds in this work are noted: eugenol, threshold ≈0.71–470 µg/Kg; linalool, threshold ≈140–600 µg/Kg; β-pinene, ≈6–1500 µg/Kg; caryophyllene, ≈64–1540 µg/Kg; 3-methylbutanal, threshold ≈0.35–7 µg/Kg; and benzaldehyde, threshold ≈300–2200 µg/Kg [46].
Importantly, it is expected that the sensory perception of volatiles is also strongly modulated by the beverage matrix. For example, the high starch content from nixtamalized corn or soaked rice, along with proteins from cocoa and broad beans, can bind hydrophobic volatiles through hydrogen bonding and hydrophobic interactions [17,22]. Additionally, the lipid fraction of cocoa butter can act as a solvent for non-polar compounds, prolonging their retention and modifying their release rate during consumption [36]. Furthermore, Maillard-derived compounds formed during cocoa roasting may interact with aroma-active molecules, influencing not only flavor perception but also aroma stability during storage [47]. This matrix effect highlights the importance of integrating both chemical and sensory data when evaluating the flavor potential of complex, traditional beverages.
On the other hand, given the complexity of the volatiles profiles of Pozol, Chilate, and Cacahuatole, a principal component analysis (PCA) was performed. It showed high variability, with PC1 + PC2/PC1 + PC3 explaining less than 30% and PC1 + PC2 + PC3 explaining less than 50% of total variance (Figure 3A–C). Nonetheless, hierarchical clustering (Figure 3D) separated the samples into two groups: one containing all Cacahuatole samples and another comprising Pozol and Chilate. It seems Cacahuatole’s distinct aldehyde profile, linked to broad bean lipid oxidation [48], contributed to this separation. The clustering also showed some samples (PZ4, CA2, CH5) diverged from their groups due to unique volatile signatures, consistent with their Venn diagram patterns, and attributed to differences in raw materials and methods used.

3.3. Sensory Evaluation of Milks with Flavors of Traditional Mexican Beverages

The processing methods, composition, volatile compound profiles, and ingredient lists were used to develop artificial flavors for each beverage in collaboration with commercial flavor houses. These flavors were then added to plain milk at the recommended doses. A trained sensory panel, familiarized with the traditional beverages, evaluated the similarity between the developed flavors and their authentic counterparts. Control samples for sensory analysis were prepared by mixing plain milk with dough samples of the traditional beverages and adding sugar. The Check-All-That-Apply (CATA) method was used to assess sensory attributes.
Figure 4 presents the CATA analysis map for the flavor proposals from commercial flavor houses (Supplementary Material, Table S1), aiming to replicate the sensory perception of traditional Mexican beverages. The panel described Pozol as having flavor notes of vanilla, sweetness, cinnamon, coffee, toasted cocoa, cooked corn, burnt corn, and an earthy taste (Supplementary Material, Table S3). Figure 4A shows that the Pozol control (FPZ0), located in the upper left quadrant, was close to descriptors associated with “optimal” flavors. As expected, the milks containing Pozol dough were perceived as having a flavor profile similar to the original beverage. FPZ0 was distant from all the flavor proposals. To the right of the y-axis, descriptors indicating the absence of key Pozol flavor notes, such as vanilla and cinnamon, as well as “unpleasant to the palate,” were found. In this area, the proposals FPZ1 and FPZ2 were located. FPZ3 and FPZ5 appeared in the lower right quadrant, near descriptors such as “light” flavors, “no cooked corn taste,” and “no toasted cocoa flavor,” highlighting their lack of essential Pozol notes. The proposal closest to the control, FPZ4, positioned in the lower left quadrant, was evaluated as “pleasant to the palate” and featured “optimal” notes of coffee, sweetness, and cinnamon.
Figure 4B displays the CATA map for Chilate-flavored samples. The panel characterized Chilate with notes of sweetness, cinnamon, vanilla, sourness, bitterness, rice, cocoa, and toasted cocoa (Supplementary Material, Table S3). Only three proposals were evaluated. The control (FCH0) was positioned near descriptors of “optimal” notes, including sweet, cocoa, and vanilla, along with slight acidity, bitterness, and rice taste. Thus, the control sample was perceived as the most similar to traditional Chilate. Proposals FCH2 and FCH3 clustered together, associated with the absence of key flavors like bitterness, roasted cocoa, and sweetness. Some descriptors indicating a slight presence of cinnamon were also nearby. In contrast, proposal FCH1 was located near descriptors such as “no vanilla, no cocoa, and no cinnamon taste” and “unpleasant to the palate.” This suggests that the developed flavors or their dosages failed to replicate the complexity of Chilate.
For Cacahuatole-flavored samples, Figure 4C illustrates the CATA analysis map, which shows similarly unsatisfactory results. The panel described Cacahuatole as a sweet drink with notes of cinnamon, vanilla, anise, roasted beans, cocoa, toasted cocoa, bitterness, and corn (Supplementary Material, Table S3). As in previous cases, the control (FCA0) was positioned near “optimal” descriptors, indicating it closely resembled the original beverage. FCA3 and FCA4, grouped in the lower right quadrant, lacked key notes such as anise, cinnamon, toasted cocoa, and corn. In the upper right quadrant, FCA1 was associated with “excessive” notes and was also perceived as “unpleasant.” Finally, FCA2 was near the center of the map, surrounded by descriptors indicating a “slight” presence of important flavors such as cinnamon, broad bean, and anise, along with “slightly similar to Cacahuatole”.
Overall, the results indicate that replicating the complex flavors of traditional beverages using artificial formulations remains challenging, owing mainly to the intricate flavor profiles of these drinks. While flavors from original Mexican beverages are mixtures with hundreds of volatiles that synergistically influence their aroma, artificial formulations generally concentrate just a few of them. Moreover, the reduced sensory fidelity of the flavored milks may also result from milk matrix effects. Milk proteins and lipids can bind hydrophobic aroma compounds, decreasing their volatility and sensory perception. Similarly, polar compounds may bind to casein micelles, slowing their release. Differences between authentic and formulated flavors appear to be due to the absence or sub-threshold levels of key volatiles and flavor suppression due to milk–volatile interactions. Further research is necessary to refine flavor development and optimize dosages to improve consumer perception since the complexity of traditional Mexican beverage aromas exceeds the capabilities of the current formulations. While this study represents an initial step in this area, deeper analysis and advanced technological development are required to achieve a more accurate replication of these flavors. This will allow the diversification of the options to taste highly culturally relevant Mexican flavors and enable their integration into milk and other dairy-based matrices, enhancing product diversity while preserving traditional sensory heritage.

4. Conclusions

Traditional Mexican cocoa-based beverages, including Pozol, Chilate, and Cacahuatole, exhibit compositions and preparation methods that impact their nutritional profiles and sensory characteristics. The volatile compound analysis revealed a diverse range of molecules influenced by ingredient inclusion and processing techniques. Terpenes, aldehydes, ketones, pyrazines, and phenolics played a crucial role in defining the sensory profiles of these beverages. Additionally, roasting cocoa beans enhanced their characteristic flavors, leading to the formation of complex aroma compounds. Future research should include quantitative analyses and compare them with odor thresholds to better understand the contribution of these volatile compounds to beverage aroma.
Despite attempts to replicate the traditional beverage flavors and their addition to milk-based formulations, commercial flavor proposals struggled to capture the sensory attributes of traditional beverages. Sensory analysis highlighted the challenge of artificially reproducing the balance of roasted notes inherent in these drinks. This underscores the need for further research into refining flavor replication to preserve the authenticity of these traditional beverages. Ultimately, the study emphasizes the cultural and nutritional significance of these drinks, warranting continued exploration into flavor development and their use in food products that improve the nutritional support for the Mexican population.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/beverages11050130/s1, Table S1. Flavor proposals for milks. Table S2. Descriptors for traditional Mexican beverages. Table S3. Phrases to describe traditional Mexican beverages. Table S4A. Volatile compound profile of Pozol. Table S4B. Volatile compound profile of Chilate. Table S4C. Volatile compound profile of Cacahuatole [49,50,51,52,53,54,55,56,57,58,59,60,61,62,63,64,65,66,67,68,69,70,71,72,73,74,75,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97,98,99,100,101,102,103,104,105,106,107,108,109,110,111,112,113,114].

Author Contributions

L.A.-G.: Conceptualization, Investigation, Acquisition of data, Formal analysis, Writing-Original draft; E.A.-P.: Investigation, Visualization, Acquisition of data, Formal analysis; P.A.V.-L.: Supervision, Data curation, Writing-review and editing; M.d.C.C.-T.: Visualization, Formal Analysis, Writing-review and editing; S.L.A.-L.: Project administration, Resources, Supervision, Writing-review and editing. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by Ganaderos Productores de Leche Pura S.A. de C.V. (ALPURA, México), SUV/DVS-EXT-2021-003.

Institutional Review Board Statement

This work was approved by the Ethics Committee of the Facultad de Química, Universidad Autónoma de Querétaro (México) CBQ20/086.

Informed Consent Statement

Participants in the sensory analysis signed informed consent forms.

Data Availability Statement

Data are available from the corresponding author on reasonable request.

Conflicts of Interest

The authors declare no conflicts of interest.

Abbreviations

The following abbreviations are used in this manuscript:
PZPozol
CACacahuatole
CHChilate
FPZFlavor proposal for Pozol
FCAFlavor proposal for Cacahuatole
FCHFlavor proposal for Chilate
PCAPrincipal component analysis
CATACheck All That Apply

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Beverages 11 00130 g001
Figure 1. Traditional fabrication process of (A) Pozol, (B) Chilate, and (C) Cacahuatole.
Figure 1. Traditional fabrication process of (A) Pozol, (B) Chilate, and (C) Cacahuatole.
Beverages 11 00130 g001
Beverages 11 00130 g002
Figure 2. Venn diagrams for volatile compounds of (A) Pozol, (B) Chilate, and (C) Cacahuatole.
Figure 2. Venn diagrams for volatile compounds of (A) Pozol, (B) Chilate, and (C) Cacahuatole.
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Beverages 11 00130 g003
Figure 3. Principal component analysis (A–C) and color map dendrogram (D) for volatile compound characterization of traditional Mexican beverages. PZ, Pozol. CH, Chilate. CA, Cacahuatole. The number after the two capital letters indicates the sample number for each beverage.
Figure 3. Principal component analysis (A–C) and color map dendrogram (D) for volatile compound characterization of traditional Mexican beverages. PZ, Pozol. CH, Chilate. CA, Cacahuatole. The number after the two capital letters indicates the sample number for each beverage.
Beverages 11 00130 g003
Beverages 11 00130 g004
Figure 4. CATA analysis map of milk formulas with flavor of traditional Mexican beverages. (A) Pozol. (B) Chilate. (C) Cacahuatole. FPZ, flavor proposal for Pozol. FCH, flavor proposal for Chilate. FCA, flavor proposal for Cacahuatole.
Figure 4. CATA analysis map of milk formulas with flavor of traditional Mexican beverages. (A) Pozol. (B) Chilate. (C) Cacahuatole. FPZ, flavor proposal for Pozol. FCH, flavor proposal for Chilate. FCA, flavor proposal for Cacahuatole.
Beverages 11 00130 g004
Table 1. Chemical and nutritional composition of traditional beverages [a].
Table 1. Chemical and nutritional composition of traditional beverages [a].
PozolChilateCacahuatole
Moisture content, %67.40 ± 1.9542.99 ± 10.5161.30 ± 4.82
Protein [b], % (d.b.)11.40 ± 1.559.31 ± 0.8016.20 ± 0.63
Fat, % (d.b.)11.78 ± 4.718.81 ± 2.6610.10 ± 0.37
Ash, % (d.b.)2.05 ± 0.362.00 ± 0.442.60 ± 0.19
Carbohydrates [c], % (d.b.)72.26 ± 6.6178.28 ± 2.6270.30 ± 1.01
Crude fiber, % (d.b.)2.50 ± 0.981.57 ± 0.370.80 ± 0.14
Energy (kJ/100 g DW)1763.57 ± 68.171737.89 ± 55.141727.10 ± 5.49
Energy (kcal/100 g DW)420.96 ± 16.38421.07 ± 13.21412.30 ± 1.31
[a] Average, standard deviation of triplicate subsamples. [b] Calculated as (N × 6.25). [c] Calculated by difference.
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MDPI and ACS Style

Abadía-García, L.; Ayala-Padilla, E.; Vázquez-Landaverde, P.A.; Cortez-Trejo, M.d.C.; Amaya-Llano, S.L. Characterization of Volatile Compounds in Traditional Mexican Cocoa-Based Beverages and Their Application in Flavored Milks. Beverages 2025, 11, 130. https://doi.org/10.3390/beverages11050130

AMA Style

Abadía-García L, Ayala-Padilla E, Vázquez-Landaverde PA, Cortez-Trejo MdC, Amaya-Llano SL. Characterization of Volatile Compounds in Traditional Mexican Cocoa-Based Beverages and Their Application in Flavored Milks. Beverages. 2025; 11(5):130. https://doi.org/10.3390/beverages11050130

Chicago/Turabian Style

Abadía-García, Lucía, Edgar Ayala-Padilla, Pedro A. Vázquez-Landaverde, María del Carmen Cortez-Trejo, and Silvia L. Amaya-Llano. 2025. "Characterization of Volatile Compounds in Traditional Mexican Cocoa-Based Beverages and Their Application in Flavored Milks" Beverages 11, no. 5: 130. https://doi.org/10.3390/beverages11050130

APA Style

Abadía-García, L., Ayala-Padilla, E., Vázquez-Landaverde, P. A., Cortez-Trejo, M. d. C., & Amaya-Llano, S. L. (2025). Characterization of Volatile Compounds in Traditional Mexican Cocoa-Based Beverages and Their Application in Flavored Milks. Beverages, 11(5), 130. https://doi.org/10.3390/beverages11050130

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